Biofilm formation in long-distance water conveyance pipelines poses significant risks to water quality, particularly under varying nutrient levels and heavy metal stress. However, the impacts of pipeline material on biofilm formation dynamics under different raw water conditions remain elusive. This study investigated the effects of nutrient availability and Fe-Mn stress on biofilm development, structural stability, bacterial community composition, and the occurrence of viable but non-culturable (VBNC) bacteria. Using reactors with different nutrient conditions, we observed that increased nutrient levels promote biofilm growth but lead to greater instability, heightening the risk of secondary contamination. Notably, nutrient escalation beyond a critical threshold had a diminishing impact on biofilm community composition. Additionally, Fe-Mn stress, while initially enhancing microbial adhesion and metabolic activity, ultimately inhibited biofilm formation over time and increases the prevalence of VBNC bacteria, particularly on stainless steel (SS) surfaces. Our findings also highlighted the importance of material selection for pipelines, with polyvinyl chloride (PVC) showing reduced biofilm formation compared to SS, making it a more suitable option for transporting raw water in environments with high metal content. Dispersal limitation determined the bacterial community assembly during the biofilm formation, accounting for 64.53–90.67 % of the variability in different scenarios. These insights offer valuable guidance for managing biofilm-related issues in water distribution systems, emphasizing the need for careful control of nutrient levels and material choice to ensure water safety over long distances.
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